In this article, in order to create a cardiac patch, a biocompatible polyurethane (PU) nanofiber polymer was developed with the electrospinning method. Bone marrow mesenchymal stem cells (BM MSCs) were isolated from whole human bone marrow. Isolated BM MSC homogeneity was determined by flow cytometry using specific markers. BM MSC-PU interactions were studied with a WST-1 kit on the 3rd and 7th days. Cardiomyocyte differentiation was performed with 5-azacytidine. Cell survival and proliferation of cardiomyocyte-like cells that were cultured on nerve growth factor (NGF)-embedded PU were evaluated with the WST-1 kit on the 3rd and 7th days. NGF's effect on the cardiomyocyte-like cell proliferation was investigated using anti-p70 S6 kinase monoclonal antibody. Our data indicate that PU nanofibers provided a suitable environment for human BM MSC, and no evident cytotoxicity was observed. Seven days after seeding, NGF-embedded nanofibers proved to be more competent in cell proliferation compared with non-NGF-embedded nanofibers. Cardiomyocyte-like cells were found to adhere on the scaffolds, showing a spreading geometry and retaining viability. Increases in p70 S6 kinase activity through NGF were monitored by flow cytometry.